Liver Cholesterol Ester Research Articles

Long noncoding RNA lincsc5d regulates hepatic cholesterol synthesis by modulating sterol C5 desaturase in large yellow croaker.

Although long noncoding RNA (lncRNA) plays a vital role in cholesterol metabolism, very little information is available in fish. Thus, a 10-week feeding experiment was performed to estimate the effects of lncRNA on cholesterol metabolism in large yellow croaker fed with fish oil (FO), soybean oil (SO), olive oil (OO), and palm oil (PO) diets. Results showed that fish fed with OO and PO diets had higher liver total cholesterol (TC) and cholesterol ester (CE) contents compared with fish fed with FO diets. Analysis of the KEGG pathway showed that the steroid biosynthesis pathway was enriched in comparisons FO vs SO, FO vs OO, and FO vs PO. Meanwhile, sterol C5 desaturase (SC5D), a cholesterol synthase, was up-regulated in the steroid biosynthesis pathway. SC5D was widely expressed in all tissues examined, and the highest expression of SC5D was detected in brain. More importantly, a novel lncRNA associated with sc5d gene was identified by RNA sequencing and named as lincsc5d. The tissue distribution of lincsc5d was similar to that of sc5d. A nuclear/cytoplasmic RNA separation assay showed that lincsc5d was a nucleus-enriched lncRNA. qRT-PCR results demonstrated that lincsc5d was markedly up-regulated in the SO, OO, and PO groups. Furthermore, the results of TC content and the lincsc5d and sc5d expression in hepatocytes agreed with in vivo results. In conclusion, this study indicated that vegetable oils, especially OO and PO, increased hepatic cholesterol levels by promoting cholesterol synthesis, and lncRNA lincsc5d and sc5d might be involved in cholesterol synthesis.

Abstract 4360: ALDH2 and the TGF-β/SMAD3 adaptor, Sptbn1 suppress hepatic steatosis and obesity

Abstract Background: Previously we have demonstrated that TGF-β-deficient mutants derived from the loss of Smad3 and the Smad adaptor β2SP (Sptbn1) phenocopy a human cancer stem cell syndrome, are exquisitely sensitive to alcohol, with dysregulation DNA damage repair and have raised levels of aldehyde dehydrogenase (ALDH2). Moreover, β2SP and β2SP/Smad3 mutant mice develop multiple liver abnormalities that include steatosis, chronic hepatitis, inflammation, and cancer. ALDH2 is a key intermediate enzyme for aldehyde and alcohol metabolism. We, therefore, hypothesized that disruption of TGF-β signaling combined with ALDH2 deficiency would increase the susceptibility of liver cancer. Methods: Aldh2 knockout mice were intercrossed with Sptbn1 heterozygous knockout mice to explore the role of Aldh2 and Sptbn1 in liver steatosis and liver cancer. Conditional knockout of Sptbn1 mice was generated by the intercross of Albumin-Cre mice with mice carrying Loxp sites flanked exons of Sptbn1. Sptbn1 conditional mice were fed with high-fat diet or chow diet. Further analysis included blood lipid panel analysis and tissue profiles including western and RNA analysis, lipidomics and metabolomics analysis of liver tissues from the mutant mice and wild-type controls. Results: Interestingly, we found that Aldh2-/-Sptbn1+/- mice on a normal diet demonstrated early signs of truncal obesity. Within 6 months of age, we observed a greater than 23% increase in body weight, with predominantly abdominal fat accumulation in Aldh2-/-Sptbn1+/- mice compared to Aldh2-/- mice or wild type (WT) mice (Aldh2-/-Sptbn1+/- vs Aldh2-/- or WT, p<0.05). A marked increase in zone 3 hepatic micro and macro-steatosis with some inflammation was observed in liver tissues of mutant Aldh2-/-Sptbn1+/- mice as well as the conditional Sptbn1 knockout (Alb-Cre+Sptbn1F/+) mice in a high-fat diet. In addition to significant increased blood glucose levels in Aldh2-/-Sptbn1+/- mice compared with Aldh2-/- mice (165±23 VS 135±13, p<0.05), further analysis revealed that the fatty liver phenotype accompanied with abnormal expression of genes related to glucose uptake, metabolisms and homeostasis as demonstrated by decrease of glucose transporter and increased expression of Ceramide which is known to promote insulin resistance. Moreover, we also observed disruption of lipid metabolism as demonstrated by significantly increased serum and liver triglyceride and cholesterol ester levels in Aldh2-/-Sptbn1+/- mice. Conclusions: Our data suggest that the TGF-β pathway plays an important role in maintaining normal glucose metabolism homeostasis and patients with SPTBN1 and ALDH2 defects may have increased susceptibility to obesity as well as fatty liver (NASH), and cancer. Citation Format: Shuyun Rao, Sobia Zaidi, Wilma Jogunoori, Shoujun Gu, Jon White, Bibhuti Mishra, Chu-Xia Deng, Patricia Latham, Lopa Mishra. ALDH2 and the TGF-β/SMAD3 adaptor, Sptbn1 suppress hepatic steatosis and obesity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4360.

Effect of Oat and Tartary Buckwheat - Based Food on Cholesterol - Lowering and Gut Microbiota in Hypercholesterolemic Hamsters.

The nutritional components in oat and tartary buckwheat had been assessed to have cholesterollowering effects. However, The effect of oat and tartary buckwheat based-food (OF) on cholesterol-lowering and gut microbiota in hypercholesterole hamsters was still limited studied because they are usually consumed in whole gran as well as after being processed. In this study, normal diets, high fat diet (HFD) with/without OF were fed to hamsters for 30 days respectively and growth parameters, metabolic parameters, and gut microbiota were investigated, respectively. It was found that OF significantly decreased plasma total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-cholesterol), lowered liver TC, cholesterol ester (CE), and triglycerides (TG) concentrations, and increased fecal weight and bile acids (BA) concentrations, compared with HFD (p < 0.05). Moreover, the concentrations of acetate, propionate, butyrate and total short-chain fatty acids (SCFAs) were significantly increased in hamsters fed with OF, compared with HFD (p < 0.05). OF changed the overall structure of gut microbiota. The relative abundances of Erysipelotrichaceae, Ruminococcaceae, and Lachnospiraceae were decreased and the relative abundance of Eubacteriaceae was increased, compared with HFD. These results suggested that OF could reduce the concentrations of plasma lipid by inhibiting cholesterol absorption in liver and promoting excretions of fecal lipid and BA. And it also increased SCFAs and modulated the gut microbiota effectively to exert the hypocholesterolemic effects.

Isolated exopolysaccharides from Lactobacillus rhamnosus GG alleviated adipogenesis mediated by TLR2 in mice.

The fibroblast cell line of 3T3-L1 was used as a cell model for screening and evaluating the feasibility of probiotic components in improving animal lipid metabolisms. The extracts from 12 Lactobacillus strains caused significantly reduced triacylglycerol (TAG) accumulation but with severe inflammation induction in 3T3-L1 adipocytes. Interestingly, exopolysaccharides (EPS) from LGG (Lactobacillus rhamnosus GG) significantly decreased the TAG accumulation without any inflammation. The anti-obesity effect of EPS was confirmed in high-fat-diets feeding mice. Fat pads of mice injected with EPS (50 mg/kg) every two days for two weeks were significantly reduced with much smaller adipocytes, compared with the counterparts. The levels of TAG and cholesterol ester in liver, as well as serum TAG, were decreased in EPS injected mice. In addition, down-regulated inflammation was observed in adipose tissue and liver. Interestingly, the expression of TLR2 in adipose tissue and 3T3-L1 cells was significantly increased by EPS addition. Moreover, the reverse of TAG accumulation in TLR2 knockdown 3T3-L1 in the presence of EPS confirmed that the inhibition effect of EPS on adipogenesis was mediated by TLR2. EPS from LGG has the potential for therapeutic development to intervene lipid metabolic disorders in mammals.

IPLA2β deficiency attenuates obesity and hepatic steatosis in ob/ob mice through hepatic fatty-acyl phospholipid remodeling

PLA2G6 or GVIA calcium-independent PLA2 (iPLA2β) is identified as one of the NAFLD modifier genes in humans, and thought to be a target for NAFLD therapy. iPLA2β is known to play a house-keeping role in phospholipid metabolism and remodeling. However, its role in NAFLD pathogenesis has not been supported by results obtained from high-fat feeding of iPLA2β–null (PKO) mice. Unlike livers of human NAFLD and genetically obese rodents, fatty liver induced by high-fat diet is not associated with depletion of hepatic phospholipids. We therefore tested whether iPLA2β could regulate obesity and hepatic steatosis in leptin-deficient mice by cross-breeding PKO with ob/ob mice to generate ob/ob–PKO mice. Here we observed an improvement in ob/ob–PKO mice with significant reduction in serum enzymes, lipids, glucose, insulin as well as improved glucose tolerance, and reduction in islet hyperplasia. The improvement in hepatic steatosis measured by liver triglycerides, fatty acids and cholesterol esters was associated with decreased expression of PPARγ and de novo lipogenesis genes, and the reversal of β-oxidation gene expression. Notably, ob/ob livers contained depleted levels of lysophospholipids and phospholipids, and iPLA2β deficiency in ob/ob–PKO livers lowers the former, but replenished the latter particularly phosphatidylethanolamine (PE) and phosphatidylcholine (PC) that contained arachidonic (AA) and docosahexaenoic (DHA) acids. Compared with WT livers, PKO livers also contained increased PE and PC containing AA and DHA. Thus, iPLA2β deficiency protected against obesity and ob/ob fatty liver which was associated with hepatic fatty-acyl phospholipid remodeling. Our results support the deleterious role of iPLA2β in severe obesity associated NAFLD.

Changes in liver lipidomics associated with sodium cholate-induced liver injury and its prevention by boiogito, a Japanese herbal medicine, in mice

Aim Bile acids play a crucial role in the development of cholestatic liver disease by mediating parenchymal cell injury and inflammation in the liver. Bile acids can also modulate lipid metabolism, although their role in the pathogenesis of cholestatic liver diseases is mostly unknown. We examined the effects of boiogito (BOT), a Japanese herbal medicine, on liver injury and lipid profile in liver lipid fractions in a mouse model of sodium cholate (CA)-induced cholestasis. Methods Lipidomics was carried out to determine the fatty acid composition in liver triglycerides, cholesterol esters, and total phospholipids using gas–liquid chromatography. Results CA induced liver injury and differential changes in fatty acid profile in triglycerides, cholesterol esters, and total phospholipids in the liver. The co-administration of BOT and CA attenuated liver injury and the changes in fatty acid profile in the liver triglycerides but not those in the liver cholesterol esters and total phospholipids. Conclusion Lipidomics indicated a unique property of liver triglyceride fatty acids in association with the preventive effect of BOT against CA-induced liver injury in mice. BOT plays an important role in hepatic TG metabolism via the prevention of cholestasis.

Effects of Dietary Corn Gluten Meal on Growth Performance and Cholesterol Metabolism in Juvenile Snakehead (Ophiocephalus argus) Fed Practical Diets

An 8-week growth trial was conducted to investigate the effects of dietary corn gluten meal (CGM) levels on growth performance and cholesterol metabolism of juvenile snakehead (Ophiocephalus argus). Six isonitrogenous (crude protein 47%) and isolipidic (crude lipid 11.5%) practical diets were formulated by replacing 0 (D1, control), 6.2 (D2), 12.3 (D3), 18.5 (D4), 22.2 (D5), and 28.4% (D6) fish meal (FM) protein with CGM protein. No significant difference in survival rate was found among dietary treatments (P&gt;0.05). No significant differences were observed in feed intake (FI), final body weight (FBW), weight gain rate (WGR), feed efficiency rate (FER) and protein productive value (PPV) among fish fed D1-D3 (P&gt;0.05). However, these indices significantly decreased with increasing CGM protein from 18.5%(D4) to 28.4%(D6) of diet (P&lt;0.05). Whole-body lipid content of fish fed D6 (28.4 %) was significantly lower than that of fish fed D1 (control) (P&lt;0.05). The plasma and liver total cholesterol, free cholesterol, cholesterol esters, LDL-C, and HDL-C levels were affected by dietary CGM protein levels. Broken line analysis based on WGR or PPV indicated that the maximum CGM protein level for the optimal growth of juvenile snakehead was 12.06% or 12.56% respectively.

H1-antihistamines exacerbate high-fat diet-induced hepatic steatosis in wild-type but not in apolipoprotein E knockout mice.

We examined the effects of two over-the-counter H1-antihistamines on the progression of fatty liver disease in male C57Bl/6 wild-type and apolipoprotein E (ApoE)-/- mice. Mice were fed a high-fat diet (HFD) for 3 mo, together with administration of either cetirizine (4 mg/kg body wt) or fexofenadine (40 mg/kg body wt) in drinking water. Antihistamine treatments increased body weight gain, gonadal fat deposition, liver weight, and hepatic steatosis in wild-type mice but not in ApoE-/- mice. Lobular inflammation, acute inflammation, and necrosis were not affected by H1-antihistamines in either genotype. Serum biomarkers of liver injury tended to increase in antihistamine-treated wild-type mice. Serum level of glucose was increased by fexofenadine, whereas lipase was increased by cetirizine. H1-antihistamines reduced the mRNA expression of ApoE and carbohydrate response element-binding protein in wild-type mice, without altering the mRNA expression of sterol regulatory element-binding protein 1c, fatty acid synthase, or ApoB100, in either genotype. Fexofenadine increased both triglycerides and cholesterol ester, whereas cetirizine increased only cholesterol ester in liver, with a concomitant decrease in serum triglycerides by both antihistamines in wild-type mice. Antihistamines increased hepatic levels of conjugated bile acids in wild-type mice, with the effect being significant in fexofenadine-treated animals. The increase was associated with changes in the expression of organic anion transport polypeptide 1b2 and bile salt export pump. These results suggest that H1-antihistamines increase the progression of fatty liver disease in wild-type mice, and there seems to be an association between the severity of disease, presence of ApoE, and increase in hepatic bile acid levels.

Bile acid and sterol metabolism with combined HMG-CoA reductase and PCSK9 suppression

Proprotein convertase subtilisin-kexin-9 (PCSK9) inhibition markedly augments the LDL lowering action of statins. The combination is being evaluated for long-term effects on atherosclerotic disease outcomes. However, effects of combined treatment on hepatic cholesterol and bile acid metabolism have not yet been reported. To study this, PCSK9-Y119X mutant (knockout) and wild-type mice were treated with or without atorvastatin for 12 weeks. Atorvastatin progressively lowered plasma LDL in each group, but no differences in liver cholesterol, cholesterol ester, or total bile acid concentrations, or in plasma total bile acid levels were seen. In contrast, atorvastatin increased fecal total bile acids (≈ 2-fold, P < 0.01) and cholesterol concentrations (≈ 3-fold, P < 0.01) versus controls for both PCSK9-Y119X and wild-type mice. All 14 individual bile acids resolved by LC-MS, including primary, secondary, and conjugated species, reflected similar increases. Expression of key liver bile acid synthesis genes CYP7A1 and CYP8B1 were ≈ 2.5-fold higher with atorvastatin in both strains, but mRNA for liver bile acid export and reuptake transporters and conjugating enzymes were not unaffected. The data suggest that hepatocyte cholesterol and bile acid homeostasis is maintained with combined PCSK9 and HMG-CoA reductase inhibition through efficient liver enzymatic conversion of LDL-derived cholesterol into bile acids and excretion of both, with undisturbed enterohepatic recycling.

Oat oil lowers the plasma and liver cholesterol concentrations by promoting the excretion of faecal lipids in hypercholesterolemic rats

The aim of the present study is to investigate the hypocholesterolemic effect of dietary oat oil in rats, fed with a hypercholesterolemic diet. The rats were divided into three groups and fed with the experiment diets for 30days, containing soybean oil, oat oil, or rice bran oil at a dose of 70g/kg. It was found that the oat oil lowered plasma total and LDL-cholesterol, and also reduced liver total, free cholesterol, cholesterol ester, and triglycerides concentrations significantly, as well as rice bran oil. Moreover, the faecal weight, total lipids, and bile acids concentrations, in the oat oil and rice bran oil groups, were significantly increased compared with that in the soybean oil group. We can conclude that dietary oat oil improves hypercholesterolemia in rats fed a hypercholesterolemic diet, by promoting excretions of faecal lipids and bile acids.

T-10, c-12 CLA Dietary Supplementation Inhibits Atherosclerotic Lesion Development Despite Adverse Cardiovascular and Hepatic Metabolic Marker Profiles

Animal and human studies have indicated that fatty acids such as the conjugated linoleic acids (CLA) found in milk could potentially alter the risk of developing metabolic disorders including diabetes and cardiovascular disease (CVD). Using susceptible rodent models (apoE−/− and LDLr−/− mice) we investigated the interrelationship between mouse strain, dietary conjugated linoleic acids and metabolic markers of CVD. Despite an adverse metabolic risk profile, atherosclerosis (measured directly by lesion area), was significantly reduced with t-10, c-12 CLA and mixed isomer CLA (Mix) supplementation in both apoE−/− (p<0.05, n = 11) and LDLr−/− mice (p<0.01, n = 10). Principal component analysis was utilized to delineate the influence of multiple plasma and tissue metabolites on the development of atherosclerosis. Group clustering by dietary supplementation was evident, with the t-10, c-12 CLA supplemented animals having distinct patterns, suggestive of hepatic insulin resistance, regardless of mouse strain. The effect of CLA supplementation on hepatic lipid and fatty acid composition was explored in the LDLr−/− strain. Dietary supplementation with t-10, c-12 CLA significantly increased liver weight (p<0.05, n = 10), triglyceride (p<0.01, n = 10) and cholesterol ester content (p<0.01, n = 10). Furthermore, t-10, c-12 CLA also increased the ratio of 18∶1 to 18∶0 fatty acid in the liver suggesting an increase in the activity of stearoyl-CoA desaturase. Changes in plasma adiponectin and liver weight with t-10, c-12 CLA supplementation were evident within 3 weeks of initiation of the diet. These observations provide evidence that the individual CLA isomers have divergent mechanisms of action and that t-10, c-12 CLA rapidly changes plasma and liver markers of metabolic syndrome, despite evidence of reduction in atherosclerosis.

Fish oil and indomethacin in combination potently reduce dyslipidemia and hepatic steatosis in LDLR −/− mice

Fish oil (FO) is a potent anti-inflammatory and lipid-lowering agent. Because inflammation can modulate lipid metabolism and vice versa, we hypothesized that combining FO with cyclooxygenase inhibitors (COXIBs), well-known anti-inflammatory drugs, can enhance the anti-inflammatory and lipid-lowering effect of FO. LDLR(-/-) mice were fed a high-fat diet supplemented with 6% olive oil or FO for 12 wk in the presence or absence of indomethacin (Indo, 6 mg/l drinking water). FO reduced plasma total cholesterol by 30% but, in combination with Indo, exerted a greater decrease (44%). The reduction of liver cholesterol ester (CE) and triglycerides (TG) by FO (63% and 41%, respectively) was enhanced by Indo (80% in CE and 64% in TG). FO + Indo greatly increased the expression of genes modulating lipid metabolism and reduced the expression of inflammatory genes compared with control. The mRNA and/or protein expression of pregnane X receptor (PXR) and cytochrome P450 isoforms that alter inflammation and/or lipid metabolism are increased to a greater extent in mice that received FO + Indo. Moroever, the nuclear level of PXR is significantly increased in FO + Indo group. Combining FO with COXIBs may exert their beneficial effects on inflammation and lipid metabolism via PXR and cytochrome P450.

The thienotriazolodiazepine Ro 11-1464 increases plasma apoA-I and promotes reverse cholesterol transport in human apoA-I transgenic mice

Ro 11-1464 is a thienotriazolodiazepine previously described to selectively stimulate apolipoprotein A-I (apoA-I) production and mRNA level in human liver cells. Here, we studied its effects upon oral administration to human apoA-I transgenic (hapoA-I) mice. HapoA-I mice were treated for 5 days with increasing doses of Ro 11-1464. Macrophage reverse cholesterol transport (mph-RCT) was assessed by following [(3) H]-cholesterol mobilization from pre-labelled i.p. injected J774 macrophages to plasma, liver and faeces. Effects on plasma lipids, apoproteins, lecithin-cholesterol : acyltransferase (LCAT) and liver enzymes, as well as on faecal excretion of cholesterol and bile salts, and on liver lipids and mRNA contents were determined. Treatment with Ro 11-1464 300 mg·kg(-1) ·day(-1) resulted in a nearly 2-fold increase in plasma apoA-I, a 2- to 3-fold increase in the level of large sized-pre-β high-density lipoprotein and a 3-fold selective up-regulation of hepatic apoA-I mRNA, but a marked decrease in all plasma lipids and LCAT activity. Mpm-RCT was decreased in blood but markedly increased in faecal sterols (4-fold) and bile acids (1.7-fold). However, liver weight and liver enzymes in plasma were also increased, in parallel with an increase in liver cholesterol ester content (all these effect being significant). In this model Ro 11-1464 causes increased hepatic expression and plasma levels of apoA-I and a suppression of LCAT, and a marked enhancement of reverse cholesterol transport, but also some symptoms of liver toxicity. The compound may therefore be a prototype for a next generation of anti-atherosclerotic medicines.

Effects of dietary cholesterol on growth performance, feed intake and cholesterol metabolism in juvenile turbot ( Scophthalmus maximus L.) fed high plant protein diets

A 9-week growth trial was conducted to investigate the effects of dietary cholesterol supplementation on growth performance, feed intake and cholesterol metabolism of juvenile turbot ( Scophthalmus maximus L.) fed high plant protein diets. A fish meal diet (diet FM) with 58% FM was formulated, and this diet was used as control. The other four diets were formulated to contain 14.5% FM, 42.0% soybean meal (SBM), and 18.5% wheat gluten meal. The four diets were supplemented with 0.0%, 0.5%, 1.0% and 1.5% cholesterol, respectively, and were isonitrogenous and isolipidic to the diet FM. They were named as diet C-0.0%, C-0.5%, C-1.0% and C-1.5%, respectively. The final dietary cholesterol concentrations were 0.30%, 0.77%, 1.25%, and 1.78%, respectively. That in control diet was 0.63%. The results showed that weight gain rate (WGR) and feed efficiency rate in fish fed diet FM were significantly higher than those in fish fed other diets ( P < 0.05). Furthermore, compared with fish fed diet C-0.0%, fish fed diet C-1.0% significantly enhanced WGR, feed intake (FI) and cholesterol levels in plasma and liver. However, WGR and FI in fish fed diet C-1.5% were significantly lower than those in fish fed diet C-1.0% ( P < 0.05). Fish fed diet C-1.0% showed significantly higher whole-body lipid content than that of fish fed other diets ( P < 0.05). The total cholesterol (TC), free cholesterol (FC), cholesterol esters, high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) contents in fish plasma, TC and cholesterol esters in fish liver, and TC in fish feces were significantly correlated with dietary cholesterol contents, and correlated coefficients were above 0.74. Fish fed diet C-0.0% showed the lowest fecal total bile acid and activity of cholesterol 7α hydroxylase among dietary treatments. These results suggested that 1.25% of dietary cholesterol is helpful for juvenile turbot fed high plant protein diets to get significantly better growth rate without negative effects.

Effects of different levels of plant proteins on the ongrowing of meagre (Argyrosomus regius) juveniles at low temperatures

Four experimental diets with different inclusion levels of plant proteins and fish protein hydrolysates were compared with a commercial diet for meagre (Argyrosomus regius) ongrowing at optimal and suboptimal water temperature. Results in terms of growth in length and weight, conversion efficiency, dietary feed intake and utilization, body composition (whole fish and liver) as well as enzyme and immunological activities are presented. Fish growth was significantly reduced by the inclusion of plant proteins, although further addition of fish protein hydrolysates improved the results. Daily feed intake was not affected by plant protein inclusion in the diets, although the group fed the highest inclusion level showed lower ingestion than the rest of the groups, probably as a consequence of a reduced dietary palatability. The decrease in water temperature during the second part of the experiment had a negative effect on feed intake and fish growth. Gross visceral morphology of meagre fed the experimental diets was not affected, but muscle weight was significantly reduced. Whole body and liver composition was not affected with plant protein inclusion. However, the inclusion of fish protein hydrolysates resulted in a significant increase in fat content, especially in liver cholesterol and steryl esters, with a parallel reduction in protein. Brush border enzymes were affected by plant protein inclusion as well as serum lysozyme that significantly increased in the fish fed the highest inclusion level. As a conclusion, up to 315 g kg⁻¹ plant protein (76.2% of total protein content) can be included in the diet for meagre without affecting growth or feed utilization. Higher inclusion levels can also be used if at least 5% fish protein hydrolysate is also included.

Hepatic One Carbon Metabolism in Diet‐Induced Obesity

Obesity is a major cause of fatty liver disease. We hypothesize that disturbances in liver one carbon metabolism contribute to the pathology of obesity‐related fatty liver disease. We studied the effect of diet‐induced obesity in mice heterozygous for targeted deletion of cystathionine‐β‐synthase (+/−) and C57BL/6J (+/+) mice fed a high fat (60% energy) diet (HF) or a control diet (C) from weaning for 12 weeks. HF+/− and HF+/+ mice had higher body weight (P&lt;0.001), greater fat mass accumulation (P&lt;0.001), higher liver triglycerides (P&lt;0.05), and lower liver cholesterol ester (P&lt;0.001) than C+/− and C+/+ mice, respectively. Independent of diet, C+/− mice had lower liver S‐adenosylmethionine (AdoMet) (P&lt;0.05) levels, and higher liver S‐adenosylhomocysteine (AdoHcy) (P&lt;0.001) levels. Both +/− and +/+ mice fed the HF diet had lower levels of liver AdoHcy (P=0.001) than mice fed the C diet. HF+/− mice had lower liver AdoMet levels (P=0.01) than C+/− mice, however this effect was not observed in +/+ mice. These findings suggest that disturbances in liver one carbon metabolism are associated with hepatic lipid accumulation in mice with diet‐induced obesity.

The effect of ghee (clarified butter) on serum lipid levels and microsomal lipid peroxidation

Ghee, also known as clarified butter, has been utilized for thousands of years in Ayurveda as a therapeutic agent. In ancient India, ghee was the preferred cooking oil. In the last several decades, ghee has been implicated in the increased prevalence of coronary artery disease (CAD) in Asian Indians due to its content of saturated fatty acids and cholesterol and, in heated ghee, cholesterol oxidation products. Our previous research on Sprague-Dawley outbred rats, which serve as a model for the general population, showed no effect of 5 and 10% ghee-supplemented diets on serum cholesterol and triglycerides. However, in Fischer inbred rats, which serve as a model for genetic predisposition to diseases, results of our previous research showed an increase in serum total cholesterol and triglyceride levels when fed a 10% ghee-supplemented diet. In the present study, we investigated the effect of 10% dietary ghee on microsomal lipid peroxidation, as well as serum lipid levels in Fischer inbred rats to assess the effect of ghee on free radical mediated processes that are implicated in many chronic diseases including cardiovascular disease. Results showed that 10% dietary ghee fed for 4 weeks did not have any significant effect on levels of serum total cholesterol, but did increase triglyceride levels in Fischer inbred rats. Ghee at a level of 10% in the diet did not increase liver microsomal lipid peroxidation or liver microsomal lipid peroxide levels. Animal studies have demonstrated many beneficial effects of ghee, including dose-dependent decreases in serum total cholesterol, low density lipoprotein (LDL), very low density lipoprotein (VLDL), and triglycerides; decreased liver total cholesterol, triglycerides, and cholesterol esters; and a lower level of nonenzymatic-induced lipid peroxidation in liver homogenate. Similar results were seen with heated (oxidized) ghee which contains cholesterol oxidation products. A preliminary clinical study showed that high doses of medicated ghee decreased serum cholesterol, triglycerides, phospholipids, and cholesterol esters in psoriasis patients. A study on a rural population in India revealed a significantly lower prevalence of coronary heart disease in men who consumed higher amounts of ghee. Research on Maharishi Amrit Kalash-4 (MAK-4), an Ayurvedic herbal mixture containing ghee, showed no effect on levels of serum cholesterol, high density lipoprotein (HDL), LDL, or triglycerides in hyperlipidemic patients who ingested MAK-4 for 18 weeks. MAK-4 inhibited the oxidation of LDL in these patients. The data available in the literature do not support a conclusion of harmful effects of the moderate consumption of ghee in the general population. Factors that may be involved in the rise of CAD in Asian Indians include the increased use of vanaspati (vegetable ghee) which contains 40% trans fatty acids, psychosocial stress, insulin resistance, and altered dietary patterns. Research findings in the literature support the beneficial effects of ghee outlined in the ancient Ayurvedic texts and the therapeutic use of ghee for thousands of years in the Ayurvedic system of medicine.

Lipid Profile of Rats Fed Cholesterol, Barely and Wheat

The comparative effects of barely and wheat on body lipids were investigated by feeding 105 growing male Sprague-Dawley rats diets containing: 1, 2, 3 and 4 g cholesterol added to 1 kg diet. Barely and wheat were added to the basal diet at 55% and the ingredients were recalculated to obtain 10% total dietary fiber. Diets were fed for 4 week; basal diet (free of cholesterol and fiber) was the control diet. Food Intake (FI); Weight Gain (WG); Total Lipid (TL) and Total Cholesterol (TC) in serum, liver, heart, kidneys spleen and feces; Triglycerides (TG), Phospholipids (PL), Free Cholesterol (FC) and Cholesterol Ester (CE) in serum and liver; Low Density Lipoprotein cholesterol (LDL-c), High Density Lipoprotein cholesterol (HDL- c) in serum and neutral sterols and bile acids in feces were studied. Rats fed barely had the lowest levels of FI, WG, TL and TC in serum and endogenous organs; TG, FC and CE in serum and liver and of LDL-c and HDL-c, whereas had the highest levels of TL, TC, neutral sterols and bile acids in feces. Added cholesterol to the diets of rats resulted in an increase, in dose dependant manner, in mentioned lipid parameters in serum and endogenous organs and its effects followed the raising order: with control diet > with wheat > with barely. However, dietary cholesterol caused an increase in TL, TC, neutral sterols and bile acids in feces.

Dietary Echium Oil Increases Tissue (n-3) Long-Chain Polyunsaturated Fatty Acids without Elevating Hepatic Lipid Concentrations in Premature Neonatal Rats ,

Echium oil (EO) contains notable quantities of both (n-6) and (n-3) PUFA and has not, to our knowledge, been studied in neonates. We compared growth, tissue PUFA concentrations, and liver lipid profiles in premature neonatal Sprague-Dawley rats that were fed an EO diet with those that were dam-fed (DF) or fed rat milk substitute (RMS) or a fish oil (FO) diet. EO or FO comprised 10% of dietary fat. Rats were delivered prematurely at d 21 of gestation by caesarean section and then DF or fed one of the diets for 6 d. Rats were killed and the fatty acid (FA) concentrations in brain, liver, ileum, and serum and liver lipid profiles were analyzed. All diet-fed rats had similar weight gain and tissue protein concentrations. Compared with DF rats, EO-fed rats had similar brain docosahexaenoic acid (DHA) levels, similar brain and liver arachidonic acid (ARA) levels, higher liver and ileal eicosapentaenoic acid (EPA) levels (P < 0.05), and similar ARA:(EPA+DHA) ratios in brain, liver, and serum. Compared with RMS-fed rats, EO-fed rats had lower liver triglyceride FA and cholesterol ester concentrations (P < 0.05), higher EPA and DHA levels in liver, ileum, and serum, a higher DHA level in brain, and lower tissue and serum ratios of total (n-6):(n-3) PUFA and ARA:(EPA + DHA) (P < 0.05). Compared with FO-fed rats, EO-fed rats had higher ARA levels in brain, liver, ileum, and serum. In conclusion, dietary EO increases tissue EPA and DHA without reducing ARA in brain and liver and without elevating hepatic lipid concentrations of premature neonatal rats.

Clofibrate-induced changes in the liver, heart, brain and white adipose lipid metabolome of Swiss-Webster mice.

Peroxisome proliferator activated receptor alpha (PPARα) agonists are anti-hyperlipidemic drugs that influence fatty acid combustion, phospholipid biosynthesis and lipoprotein metabolism. To evaluate impacts on other aspects of lipid metabolism, we applied targeted metabolomics to liver, heart, brain and white adipose tissue samples from male Swiss-Webster mice exposed to a 5 day, 500 mg/kg/day regimen of i.p. clofibrate. Tissue concentrations of free fatty acids and the fatty acid content of sphingomyelin, cardiolipin, cholesterol esters, triglycerides and phospholipids were quantified. Responses were tissue-specific, with changes observed in the liver > heart ≫ brain > adipose. These results indicate that liver saturated fatty acid-rich triglycerides feeds clofibrate-induced monounsaturated fatty acid (MUFA) synthesis, which were incorporated into hepatic phospholipids and sphingomyelin. In addition, selective enrichment of docosahexeneoic acid in the phosphatidylserine of liver (1.7-fold), heart (1.6-fold) and brain (1.5-fold) suggests a clofibrate-dependent systemic activation of phosphatidylserine synthetase 2. Furthermore, the observed ~20% decline in cardiac sphingomyelin is consistent with activation of a sphingomeylinase with a substrate preference for polyunsaturate-containing sphingomyelin. Finally, perturbations in the liver, brain, and adipose cholesterol esters were observed, with clofibrate exposure elevating brain cholesterol arachidonyl-esters ~20-fold. Thus, while supporting previous findings, this study has identified novel impacts of PPARα agonist exposure on lipid metabolism that should be further explored.